猫棒束孢乙醇提取物对荷H22肝癌小鼠的抗肿瘤作用  

Antitumor effect of alcohol extract of Isaria felina on H22 hepatoma tumor-bearing mice

作　　者：余国彦 史晓伟 张艺尹 陈丽霞 赵莉莉[2] 杨永明[2] 王靖[2] 阎磊[2] 杨喜花[1] Yu Guoyan;Shi Xiaowei;Zhang Yiyin;Chen Lixia;Zhao Lili;Yang Yongming;Wang Jing;Yan Lei;Yang Xihua(School of Public Health,Shanxi Medical University,Taiyuan 030001,China;Experimental Animal Center of Shanxi Province Cancer Hospital,Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences,Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan 030013,China)

机构地区：[1]山西医科大学公共卫生学院,太原030001 [2]山西省肿瘤医院中国医学科学院肿瘤医院山西医院山西医科大学附属肿瘤医院实验动物中心,太原030013

出　　处：《肿瘤研究与临床》2024年第4期241-247,共7页Cancer Research and Clinic

基　　金：山西省卫生健康委科研课题(2021072)。

摘　　要：目的探讨猫棒束孢乙醇提取物(IFA)对荷H22肝癌小鼠的抗肿瘤作用及可能机制。方法将猫棒束孢用50%乙醇重复提取两次,合并两次滤液,水浴蒸干,得到IFA。将36只雌性昆明小鼠按照随机数字表法分为6组,分别为正常对照组、模型对照组、阳性对照组、IFA低剂量组、IFA中剂量组、IFA高剂量组,每组6只;除正常对照组外,其余各组于小鼠右侧腋窝皮下接种小鼠肝癌H22细胞,建立荷瘤小鼠模型。正常对照组、模型对照组均每天胃灌注0.9%NaCl溶液,阳性对照组每天腹腔注射25 mg/kg环磷酰胺并胃灌注0.9%NaCl溶液,IFA低、中、高剂量组分别每天胃灌注37.5、75、150 mg/kg IFA。连续给药10 d后,麻醉后摘眼球取血,颈椎脱臼法处死小鼠,分离肿瘤组织、胸腺和脾脏,称量各标本质量;根据肿瘤组织质量和脏器质量分别计算抑瘤率和免疫器官指数;采用酶联免疫吸附试验检测血清转化生长因子β(TGF-β)水平;HE染色观察肿瘤组织细胞形态;对肿瘤组织进行TUNEL荧光染色,显微镜下观察细胞凋亡情况,并计算总细胞中凋亡细胞占比;免疫组织化学法检测肿瘤组织中bcl-2和bax蛋白表达水平,用ImageJ软件分析总面积中bcl-2或bax蛋白阳性区域占比。结果接种H22细胞6 d后,小鼠皮下接种部位有结节产生,提示造模成功。给药10 d后,阳性对照组及IFA低、中、高剂量组荷瘤小鼠的抑瘤率分别为63.94%、35.70%、62.44%、72.67%。阳性对照组小鼠胸腺指数低于模型对照组和正常对照组,差异均有统计学意义(均P<0.05);IFA各剂量组与正常对照组和与模型对照组间胸腺指数差异均无统计学意义(均P>0.05),IFA各剂量组胸腺指数均高于阳性对照组,差异均有统计学意义(均P<0.05)。各组小鼠间脾脏指数差异均无统计学意义(均P>0.05)。IFA中剂量组、IFA高剂量组小鼠血清TGF-β浓度均低于正常对照组、模型对照组、阳性对照组,差异均有统计学Objective To investigate the antitumor effect of alcohol extract of Isaria felina(IFA)on H22 hepatoma tumor-bearing mice and its possible mechanisms.Methods Isaria felina was extracted twice with 50% alcohol,combined with filtrate twice,and steamed in water bath,and then IFA was obtained.According to the random number table method,36 female Kunming mice were divided into 6 groups,including the normal control group,the model control group,the positive control group,IFA low,medium and high dose groups,with 6 mice in each group.The mice in the other 5 groups were subcutaneously inoculated with hepatoma H22 cells in the right axilla except for the normal control group,and then the tumor-bearing mice models were established.The mice in the normal control group and the model control group were injected with 0.9%NaCl solution,respectively per day.The mouse in the positive control group was intraperitoneally injected with 25 mg/kg cyclophosphamide and 0.9%NaCl solution.IFA low,medium and high dose groups were injected with 37.5,75 and 150 mg/kg IFA per day.After 10 days of continuous administration,blood was collected from the eyeball after anaesthesia,and cervical spine dislocation method was used to kill the mice;and then the tumor tissue,thymus and spleen were separated and weighted.Tumor inhibition rate and immune organ index were calculated according to tumor tissue weight and organ weight.Enzyme-linked immunosorbent assay(ELISA)was used to detect the serum level of transforming growth factor-β(TGF-β).The morphology of tumor tissues was observed by using HE staining.TUNEL method was used to stain the tumor tissues.The cell apoptosis was observed under microscope and the proportion of apoptotic cells in total cells was calculated.The expression levels of bcl-2 and bax proteins in the tumor tissues were detected by using immunohistochemistry.The ImageJ software was used to analyze the proportion of bcl-2 or bax protein positive areas in the total area.Results Nodules were observed in the subcutaneous inoculation sit

关 键 词：肝肿瘤 疾病模型 动物 猫棒束孢 治疗结果 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]